Human primary cortical neurons were prepared in our laboratory by Dr. Darbinyan [4 (link),25 (link)–28 (link)]. Human fetal brains resulting from elective abortion were obtained from Advanced Bioscience Resources (ABR), Inc., Alameda, CA 94501 USA, under a protocol approved by Temple University’s IRB. The protocol also complied with NIH guidelines at the time, although currently, NIH guidelines do not permit new collection of human fetal tissues. In brief, 16-weeks fetal brain (approx 13 g) was collected and treated with Tryple Express enzyme (Invitrogen, CA), DNase I (10 U/mL; Sigma, St. Eouis, MO) for 15 min at 7°C, then washed three times with Hibernate E medium. Neurobasal medium containing B27 supplement and 0.25 mM Glutamax was used for a tissue trituration with a glass Pasteur pipette, and cells were then plated on poly-D-lysine-coated 60-mm dishes (Sigma). After 16 hours, 1 μM of Cytosine arabinoside Ara-C (Sigma) was added to cells for 48 hours. Cells were cultured in Neurobasal medium containing several antibiotics, including 10 μg/mL gentamycin, 100 units/mE penicillin and 10 μg/mL streptomycin. Medium also contained one μg of antifungal fungizone (Life Technologies, Inc.). Cells were maintained at 37°C incubator.
Poly d lysine coated 60 mm dishes
Manufactured by Merck Group
Poly-D-lysine-coated 60-mm dishes are laboratory equipment used for cell culture applications. They provide a charged surface that promotes cell adhesion and growth.
Automatically generated - may contain errors
2 protocols using poly d lysine coated 60 mm dishes
1
Preparation of Primary Human Cortical Neurons
2
Preparation of Primary Human Cortical Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human primary cortical neurons were prepared in our laboratory by Dr. Darbinyan [4 (link),25 (link)–28 (link)]. Human fetal brains resulting from elective abortion were obtained from Advanced Bioscience Resources (ABR), Inc., Alameda, CA 94501 USA, under a protocol approved by Temple University’s IRB. The protocol also complied with NIH guidelines at the time, although currently, NIH guidelines do not permit new collection of human fetal tissues. In brief, 16-weeks fetal brain (approx 13 g) was collected and treated with Tryple Express enzyme (Invitrogen, CA), DNase I (10 U/mL; Sigma, St. Eouis, MO) for 15 min at 7°C, then washed three times with Hibernate E medium. Neurobasal medium containing B27 supplement and 0.25 mM Glutamax was used for a tissue trituration with a glass Pasteur pipette, and cells were then plated on poly-D-lysine-coated 60-mm dishes (Sigma). After 16 hours, 1 μM of Cytosine arabinoside Ara-C (Sigma) was added to cells for 48 hours. Cells were cultured in Neurobasal medium containing several antibiotics, including 10 μg/mL gentamycin, 100 units/mE penicillin and 10 μg/mL streptomycin. Medium also contained one μg of antifungal fungizone (Life Technologies, Inc.). Cells were maintained at 37°C incubator.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!